[Diet in rheumatic disease]. / Ernährung bei rheumatischen Erkrankungen.

The goals of dietary therapy in rheumatic diseases are alleviation of under- and malnutrition, inhibition of inflammation, prophylaxis of osteoporosis, as well as recognition and treatment of nutrient sensitivities or intolerances.Inflammation inhibition in these patients is improved by manipulating the omega-3/omega-6 fatty acids ratio in the diet. Reduction of dietary arachidonic acid is recommended. This polyunsaturated fatty acid is the main precursor of pro-inflammatory mediators which interact with chemokines und cytokines. Simultaneously, intake of anti-inflammatory omega-3 fatty acids is increased. Studies have shown that this dietary regimen results in an amelioration of symptoms in patients with inflammatory rheumatic diseases. Dietary therapy in rheumatic diseases is often complicated by concomitant diseases and physical handicaps necessitating interdisciplinary patient care, consisting of rheumatologists, nutritionists, physiotherapists and ergotherapists. Dietary therapy of rheumatic diseases is an adjuvant therapy that should be initiated after a patient is properly diagnosed.

Human Opisthorchis and Haplorchis infections in Laos.

A study of the epidemiology of trematode infections was carried out in Vientiane province, Laos, comprising examination of snails, fish, cats, and human stools. Opisthorchis viverrini cercariae were found in the snail Bithynia siamensis goniomphalus (Bithyniidae) and Haplorchis taichui cercariae in Tarebia granifera (Thiaridae). O. viverrini metacercariae were recorded in the flesh of 7 species of cyprinid fish; metacercariae of the heterophyid fluke Haplorchis taichui were found in 4 species of cyprinid fish and those of H. pumilio in 2 species. Cats from the same localities were infected with O. viverrini and 4 species of heterophyid flukes, frequently as multiple infections. The highest prevalence of small fluke eggs in human stools was recorded in the 20-29 years age group, particularly in males (90.4%). Three-quarters of human infections were light (less than 1000 eggs/g), the highest value (2975 eggs/g) being recorded in the 10-14 years age group. Identification of small flukes excreted by Laotian patients after treatment with praziquantel in Czechoslovakia revealed more cases with H. taichui than with O. viverrini. The results confirmed the simultaneous occurrence of O. viverrini and intestinal heterophyid flukes in the area studied.

The fate of Cryptosporidium parvum oocysts ingested by dung beetles and their possible role in the dissemination of cryptosporidiosis.

The fate of oocysts of Cryptosporidium parvum ingested by dung beetles and the possible role these beetles serve in the dissemination of cryptosporidiosis were tested on the following species: Anoplotrupes stercorosus, Aphodius rufus, and Onthophagus fracticornis. Ten specimens of each species were offered cattle dung supplemented with 5.9 x 10(6) oocysts of C. purvum. After 24 hr of feeding, the beetles were examined for the presence of oocysts on their external surfaces, in their gastrointestinal tracts, and in feces passed during the experiment. Results indicate that although many oocysts pass safely through the mouthparts and gastrointestinal tracts of the beetles, the majority of them are destroyed. Coprophagous insects can, therefore, be considered an important aspect in the ecology of gastrointestinal diseases of man and livestock, as both agents of control and dissemination.

Natural focus of microsporosis caused by Microsporum canis Bodin.

The authors discovered a natural focus of microsporosis caused by the dermatophyte Microsporum canis Bodin. An adult man and four children were afflicted by this dermatophytosis. Symptomatic and inapparent forms of the disease in the focus were also detected in several cats and dogs. Homeless cats were found to be the source of the causative agent and its permanent hosts. A chainlike transmission from these cats was triggered to home-kept cats and dogs and to family members of their owners. Both, the clinical picture of the disease and characteristics of the strains isolated corresponds with the published data. The authors summed up the evidence collected to date on the epidemiology of microsporoses caused by M. canis.

The serological surveillance of several groups of patients using antigens of Encephalitozoon hellem and E. cuniculi antibodies to microsporidia in patients.

This study was undertaken to attempt to identify correlations between microsporidial seroprevalence data in man, clinical diseases and groups of people at the risk of HIV/AIDS infection. Groups of patients were selected according to the predilection of members of the genus Encephalitozoon for nervous and kidney tissue. Female prostitutes and alcohol and intravenous drug abusers were selected as groups at risk of HIV/AIDS infections. A total of 401 samples of human sera were examined for the presence of antimicrosporidial IgG antibodies by ELISA test with a titre of 600 considered borderline positivity. The highest occurrence of antimicrosporidial antibodies was found in the groups of alcohol abusers (16% from 43 patients), intravenous drug abusers (11% from 9 patients) and prostitutes (10% from 80 women) for E. cuniculi antigen and in the groups of psychiatric patients (14% from 44 patients), malaria patients (11% from 38 patients) and alcohol abusers (7% from 43 patients) for E. hellem antigen. The occurrence of specific antibodies of the six examined diagnostic units (glomerulonephritis chronica, pyelonephritis chronica, schizophrenia, dementia, multiple sclerosis and cerebral stroke) was statistically significant only in patients with pyelonephritis chronica and dementia (p < 0.05). No cases of microsporidial infection were found among the female prostitutes by parasitological examination, although one case of giardiasis was identified. Sera of patients with high anti-E. cuniculi and anti-E. hellem antibodies (titres in ELISA of 600 and above) were confirmed by Western blot using E. cuniculi and E. hellem polypeptides, respectively. These results suggest that the examined patients could show residual antibodies from past or latent infections.

Antigenic characterization of human isolates of cryptosporidia.

Isolates of cryptosporidia from immunodeficient patients and from immunocompetent children suffering from diarrhoea were sources for preparation of antigens. Altogether, antigens from 21 isolates were obtained, 19 from children and 2 from AIDS patients. With one exception, all children were under 4 years old and most of them were between 1 and 2 years old. The probable source of infection was found in 11 cases. In 8 cases, as well as in both AIDS patients, the source of infection was not determined. The same groups of persons and the control group of healthy blood donors were examined serologically using ELISA with the antigen of bovine origin and positive sera were used for following experiments. Soluble and membrane-enriched antigens from oocysts were compared using polyacrylamide gel electrophoresis and electroimmunotransfer blots developed by human immune sera. While no differences were recorded in soluble antigens, two of the membrane-enriched antigens were missing (15.5 and 33 kD bands). While all isolates with a pattern typical for Cryptosporidium parvum were infective for neonatal mice, both isolates with missing bands were not infective for newborn mice in repeated experiments. The first of them was infective for chickens and was originally determined to be Cryptosporidium baileyi (Ditrich et al. 1991). However, the antigenic pattern differs from pattern of this species. The second isolate was infective for guinea pigs and its species classification remains uncertain.

Amoebiasis in foreign students.

A total of 2,883 foreign students at the age of 18-30 years were examined for amoebiasis after their arrival to Czechoslovakia. Stool examinations revealed the presence of Entamoeba histolytica in 112 of them (3.9%). Students from 38 countries were found to be infected with this parasite. In a set of 2,064 students from these countries E. histolytica prevalence in stool was 5.4%. There were greater differences in the prevalence between individual countries inside a geographical region than between individual geographical regions. The highest E. histolytica prevalence in stool was found in students from tropical and southern Africa (6.7% of 745 examined) and the lowest in students from South-eastern Asia (3.1% of 321 examined). In a simple cross-section study, antibodies against E. histolytica were detected by enzyme-linked immunosorbent assay (ELISA) in the sera of 1,001 persons. Antibodies were detected in 7.9% of students at the following titres: 1:200 in 4.5%, 1:600 in 1.5%, 1:1,800 in 1.9%. Antibodies occurred more frequently in students carrying E. histolytica cysts (X2 = 14.9). Titre of ELISA antibodies in patients with confirmed liver abscess was higher than 1:1,800. counterimmunoelectrophoresis (CIEP) test was used for serum examinations of patients who had been demonstrated by ELISA to be seropositive and of those carrying E. histolytica cysts. In a set of 170 patients CIEP antibodies were also more frequent in those carrying E. histolytica cysts (X2 = 26.95). A comparison of the results of ELISA and CIEP tests in the same patients revealed that CIEP antibodies were more dependent on the actual parasitization with E. histolytica than ELISA antibodies.(ABSTRACT TRUNCATED AT 250 WORDS)

Cytokine response to infection with the microsporidian, Encephalitozoon cuniculi.

The production of three cytokines, interferon gamma (IFN-gamma), interleukin 10 (IL-10) and interleukin 12 (IL-12), was measured after intraperitoneal infection of immunocompetent Balb/c mice and immunodeficient SCID mice with the microsporidian, Encephalitozoon cuniculi Levaditi, Nicolau et Schoen, 1923. High levels of IFN-gamma were detected in ex vivo cultures of peritoneal exudate cells (PEC) of Balb/c mice, a lower, but earlier IFN-gamma response was observed in PEC from SCID mice. The early IL-10 response was detected in ex vivo cultures of splenocytes from Balb/c but not from SCID mice, explaining a delay in the IFN-gamma response in Balb/c mice. IL-12 was detected in PEC cultures from SCID mice, indicating an alternative pathway of IFN-gamma production by NK cells stimulated by IL-12 derived from macrophages.

Histopathological diagnostics of liver amoebiasis in Cambodia.

Results of histopathological examination of a solitary chronical amoebic abscess in the liver of a patient from Cambodia are described. The abscess was at a proliferously reparative phase and Entamoeba histolytica was detected at the border between the necrosis and inner wall of abscess, as well as in the proper non-specific granulation tissue of the inner layers of abscess capsule. A differential diagnosis of the amoebic abscess from similar parasitary or pseudoparasitary liver lesions and differential diagnosis of amoebae in histological sections are given. For orientation examinations of the liver tissue for the presence of amoebae the authors recommend the impregnation after Grocott and staining with Goldner's trichrome for a more detailed evaluation of histological sections.

Analysis of soluble Entamoeba histolytica antigen using enzyme-L inked electroimmunotransfer blots.

Soluble antigens of ten strains of E. histolytica were studied by sodium dodecylsulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and enzyme-linked electroimmunotransfer blots (EITB). No relations of immune replicas to virulence, geographical origin and method of cultivation (xenic or axenic culture) were found. Antigens of all ten strains tested precipitated with anti-E. histolytica human serum in the area of 30-43 kD. Antigen of HK-9 strain created in this area a characteristic pattern with all sera containing the specific anti-E. histolytica antibodies and, therefore, EITB can be used for excluding false positive results in ELISA.

Gastrointestinal parasitoses discovered in agricultural workers in South Bohemia, Czechoslovakia.

In the years from 1975 to 1982 1,750 persons, mostly employed by agricultural enterprises in the South Bohemian Region in Czechoslovakia, were examined. We discovered seven species of parasites: Taenia saginata in 0.3%, Enterobius vermicularis in 10.1%, Giardia lamblia in 1.0%, Endolimax nana in 0.8%, Entamoeba coli in 0.7%, Entamoeba hartmanni in 0.2%, and Chilomastix mesnili in 0.5%. The greatest number of parasites was found in students of the Secondary agricultural and technical school. Only two species of parasites were diagnosed in children of the employees. The incidence of E. vermicularis was 75% in children, in adult employees of agricultural enterprises, however, only 9.8%.

Entamoeba polecki: morphology, immunology, antigen study and clinic of the first infections in Czechoslovakia.

Entamoeba polecki Prowazek, 1912 was recorded for the first time in Czechoslovakia in two students from Kampuchea. Uninucleated cysts of diameter 14.2-15.7 micron with nuclei diameter 3.2-4.2 micron were found-repeatedly in stool samples taken from them. The nucleus accounted in average for 24.3% of the cyst diameter. The isolation and two following subinoculations on Dobell-Leidlaw medium were achieved, more abundant growth was recorded on medium with pig serum. Sera of both students were positive in serological tests using the E. histolytica antigen. No serious clinical symptoms were observed, both patients were cured successfully by metronidazole and ornidazole. Electroimmunotransfer blots were used to characterize E. polecki as a separate species. The antigenic structure of polyxenically grown E. polecki was compared with the antigenic structure of E. histolytica (axenically grown HK-9 strain and 3 polyxenically grown strains). Different blot patterns of both species were obtained, but common fractions of 30-40 kD probably responsible for serological cross-reactions were found.

The severe combined immunodeficient mouse as a model for Encephalitozoon cuniculi microsporidiosis.

Microsporidia have been recently recognized as opportunistic pathogens in AIDS patients. In attempt to develop an animal model with features similar to the infections observed in the immunodeficient patients, the adult severe combined immunodeficient mice (SCID) were administered both intraperitoneally and perorally by 2 x 10(7) spores of the murine isolate of E. cuniculi. The experimental inoculation caused a severe, fatal disease characterized by the dissemination of microsporidia into the host tissues. The dominant route of E. cuniculi dissemination in the SCID mice was continual direct extension from the site of inoculation to adjacent tissues and organs, terminating in hematogenous spread of infection in the host. The different courses of microsporidiosis in SCID mice relative to the mode of inoculation (i.p. vs. p.o.) was observed. The survival time of i.p. infected SCID mice was 3 weeks--vs. 5 weeks in p.o. infected SCID mice. Experimental microsporidiosis in SCID mice should provide a useful model for studies in microsporidial pathogenesis, mechanisms of resistance, immunotherapy, and in evaluating potential antimicrosporidial agents.

Prophylactic and therapeutic immune reconstitution of SCID mice infected with Encephalitozoon cuniculi.

Severe combined immunodeficient (SCID) mice develop lethal infections, resembling opportunistic microsporidiosis of immunocompromised patients, after intraperitoneal (i.p.) inoculations of spores of Encephalitozoon cuniculi. In the present study, SCID mice reconstituted i.p. with 5 x 10(7) spleen cells from naive adult BALB/c mice 14 days prior to the i.p. injection of 10(7) spores were completely resistant to the infection, whereas control infected SCID mice developed clinical disease and died within 17 days post infection (DPI). In another experiment, SCID mice infected i.p. with 10(7) spores of E. cuniculi and after that (on DPI 7) injected i.p. with 5 x 10(7) spleen lymphocytes isolated from immune adult BALB/c mice were partially protected against the parasite (40% of the reconstituted mice survived). In both experiments, high levels of parasite-specific serum antibodies (mostly of the IgG-isotype) were detected in the infected immunocompetent BALB/c mice, whereas virtually no antibodies were found in the infected SCID mice. However, SCID mice reconstituted with either naive spleen cells or immune lymphocytes revealed humoral immune responses comparable with those of immunocompetent mice.

Comparative morphology of eggs of the Haplorchiinae (Trematoda: Heterophyidae) and some other medically important heterophyid and opisthorchiid flukes.

The egg morphology of the following medically important small flukes from Southeast Asia and Far East were studied: Opisthorchis viverrini, Clonorchis sinensis (Opisthorchiidae), Haplorchis taichui, H. pumilio, H. yokogawai, Stellantchasmus falcatus and Metagonimus sp. (Heterophydiae). This study revealed a great intraspecific variability and interspecific similarity in size and shape of eggs. The eggs shape does not seem to be suitable for species identification. On the other hand, biometrical analysis of egg size enabled us to divide eggs from the species studied into four distinct groups according to the Faust-Meleney index (FMI) characterizing egg size rather than the length and width of eggs. The surface structures of eggs, delineated by using a scanning electron microscope (SEM), appeared to be a suitable morphological feature for distinguishing some groups of small flukes. Eggs from the Haplorchiinae were typified by the characteristic filamentous mesh structure. The problems of identification of eggs in human stool samples and suitability of using morphological criteria such as shape and size of eggs are discussed herein.

Entamoeba histolytica--the virulence of imported strains.

We tested the virulence of 15 strains of Entamoeba histolytica, imported to Czechoslovakia, by intracaecal inoculation of laboratory rats. According to the scoring system of Neal, none of the 15 strains possessed the virulence index greater than 2. This indicates that all the organisms tested should be classified as avirulent. However, it should be noted that all the strains produced infection of the caecum and thus should be considered infective for rat. For 7 strains, isoenzyme patterns were determined for PGM, HK and ME. One imported strain, obtained from student from Congo, demonstrated isoenzyme pattern for PGM and HK indicated that the strain was virulent. This organisms had the index of virulence 1.8 (avirulent) in animal experiment; it was isolated from cysts of clinically asymptomatic patient. Examination of the rectal mucosa of the donor of the strain indicated typical chronic catarrhal proctitis of mild degree. Examination of the patient's serum demonstrated the presence of anti E. histolytica antibodies by CIEP, while the ELISA test was negative. Twenty-one cyst carriers were examined by rectoscopy. Pathologic changes were observed in 20 of these, as follows: altered vascular structure (13X), roughened mucosa (12X), mucosal reddening (10X), decreased glistening (7X), mucus in mucosa (5X), inflammatory pseudopolypes (2X), ulcers (2X), enanthema (1X). Histological biopsies were obtained in 15 cases. One was considered normal. Remaining 14 biopsies exhibited following morphological changes: increased mucus secretion (8X), edema (7X), lymphocytic and plasmocytic infiltration (6X), lymphocytic and plasmocytic infiltration in addition to the presence of eosinophilic granulocytes (6X), presence of mucophages (5X), haemorrhages (4X), increased vascularity (3X), lymphocytic and plasmocytic infiltration with presence of extremely abundant eosinophilic granulocytes (1X), erosive-ulcerative changes of mucosa (1X). The changes observed indicated chronic catarrhal proctitis with expression to greater or less degree of signs of chronic catarrhal inflammation.

Differential diagnosis of opisthorchiid and heterophyid metacercariae (Trematoda) infecting flesh of cyprinid fish from Nam Ngum Dam Lake in Laos.

The examination of flesh of 313 cyprinid fish from the Nam Ngum water reservoir in Vientiane Province, Laos, revealed the presence of four morphologically similar opisthorchiid and heterophyid metacercariae of medical importance. The following morphological characters, visible under dissecting microscope with low magnification, were typical of trematodes recorded in fish: (1) Opisthorchis viverrini, the liver fluke, has oval cysts containing quickly moving C-shaped metacercariae provided with a rather large excretory bladder; (2) metacercariae of Haplorchis pumilio and H. taichui are surrounded by a thin-walled, oval cyst; they are actively motile, C- or S-shaped, and possess a relatively small excretory bladder; (3) cysts of Haplorchoides mehrai are considerably variable in shape, size, and thickness of cyst wall; larvae are slowly moving, C- or S-shaped, or often not folded, and provided with a rather small excretory bladder. Some cysts with dead or degenerated H. mehrai larvae are filled with brownish or yellowish pigment. Morphological variability of some larvae, however, excluded correct identification of all metacercariae during routine examination of fish. Therefore, we consider isolation of several larvae from each fish host as well as all "atypical" metacercariae from fish flesh and their examination under the light microscope to be necessary. Even though this method of examination is rather time consuming, it decreases the possibility of false identification of trematode larvae encysted in flesh of cyprinid fish during field studies on opisthorchosis and heterophyiosis.

Occurrence of some medically important flukes (Trematoda: Opisthorchiidae and Heterophyidae) in Nam Ngum water reservoir, Laos.

Occurrence of some medically important flukes transmitted to man by eating of raw fish was observed at 18 localities of Nam Ngum water reservoir, an important source of fish for inhabitants of Vientiane Province, Laos. Cercariae of Opisthorchis viverrini were recorded in 0.9% of Bithynia siamensis goniomphalus examined but in the most populated sites, where the density of the snails was the highest up to 2.2% of the intermediate hosts were infected. Haplorchis cercariae were found in 2.0% of Tarebia granifera examined. O. viverrini metacercariae were found in flesh of four fish species (Hampala dispar. H. macrolepidota, Puntius gonionotus, Cyclocheilichthys repasson) from 9 localities with the highest prevalence (16%) in H. dispar. Metacercariae of Haplorchis pumilio and H. taichui were found in flesh of Hampala dispar, H. macrolepidota and Puntius gonionotus with the prevalence of infection 2-6%. Metacercarie of Stellantchasmus falcatus were localized in fins of Xenentodon canciloides. Domestic cats (Felis catus f. domestica) harbored the following trematode species: O. viverrini, H. pumilio, H. taichui, H. yokogawai, S. falcatus. Eggs of small flukes were found in stools of 44% of examined inhabitants of 3 villages and of the district center Thinkeo. The study revealed that the best predictor of the existence of small fluke foci and their species composition in the area studied was the second intermediate host-fish.

Study of the body surface of Haplorchis yokogawai (Katsuta, 1932) and H. taichui (Nishigori, 1924) (Trematoda:Heterophyidae).

The morphology of the surface of two heterophyid flukes, Haplorchis yokogawai and H. taichui, that can infect humans eating raw fish flesh, has been studied using scanning and transmission electron microscopy. The body surface of both species is covered with scale-like, serrated spines with the exception of areas around the oral sucker, ventrogenital opening and excretory pore. The species slightly differ by the dentation pattern of tegumental spines: H. yokogawai has finer and more numerous spine teeth (maximum tooth number 14-16) while H. taichui has more serrated spines, mostly with a lower number (up to 10-12) of somewhat more robust teeth. However, because of the variability, the shape and dentation of spines do not seem to be reliable diagnostic features enabling mutual differentiation of both species.

Serological differentiation of human small fluke infections using Opisthorchis viverrini and Haplorchis taichui antigens.

Sera from 642 inhabitants of Vientiane Province (Laos) were examined by enzyme-linked immunosorbent assay (ELISA) using cytoplasmic and membranous antigens prepared from adult worms. Worms of Opisthorchis viverrini originated from liver of dissected cats, Haplorchis taichui were obtained from a stool specimen of a Laotian patient after praziquantel treatment. The sera were divided into five groups according to the intensity of infection expressed as egg count per gram of patients stool (EPG). Correlation between intensity of infection and the level of antibodies in serum was recorded. Reactions obtained using the cytoplasmic antigens were more sensitive and more specific compared to those with membranous antigens. Cross-reactions between antigens of both helminth species were found. Highly positive sera were examined using electroimmunotransfer blots (EITB) with cytoplasmic antigens of both species, which enabled the species differentiation. Antigens of both species yielded several shared fractions; however, differences between them were found: homologous sera reacted specifically with O. viverrini antigen in the area of 70 kDa and with H. taichui antigen in the area of 10 kDa. Thirty-one of 122 tested sera had specific antibodies against O. viverrini, 77 sera against H. taichui and 14 sera against both species. The results confirmed our assumption about predominant occurrence of heterophyid flukes in the human population living in studied area, compared with the occurrence of opisthorchid flukes. Hence, serology seems to be helpful tool for correct diagnosis of small fluke infections.